Accelerated aneurysmal dilation associated with apoptosis and inflammation in a newly developed calcium phosphate rodent abdominal aortic aneurysm model.
Accelerated aneurysmal dilation associated with apoptosis and inflammation in a newly developed calcium phosphate rodent abdominal aortic aneurysm model.
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DOI：
10.1016/j.jvs.2012.01.038
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发表时间：
2012-08
影响因子：
4.3
通讯作者：
Liu, Bo
中科院分区：
文献类型：
--
作者：
Yamanouchi, Dai;Morgan, Stephanie;Stair, Colin;Seedial, Stephen;Lengfeld, Justin;Kent, K. Craig;Liu, Bo
氯化钙（CaCl 2）模型是广泛接受的腹主动脉瘤（AAA）啮齿动物模型。钙沉积，主要由磷酸钙（CaPO 4）晶体组成，已报告存在于人类和实验动脉瘤中。将CaCl 2和磷酸盐缓冲液（PBS）混合，将CaPO 4晶体用于体外DNA转染。在这里，我们描述了加速动脉瘤形成的CaCl 2模型的修改。通过在CaCl 2处理后将PBS施加到小鼠肾下主动脉上来创建改良的CaCl 2，即CaPO 4模型。对CaPO 4模型和常规CaCl 2模型的动脉进行形态学、组织学和免疫组织化学分析。使用CaCl 2和PBS的混合物进行体外方法以产生CaPO 4晶体。用DNA片段化ELISA法检测CaPO 4诱导原代培养的小鼠血管平滑肌细胞（VSMCs）凋亡。首先，我们证明了CaPO 4模型产生AAA，定义为主动脉直径增加50%或更大;比CaCl 2模型更快。CaPO 4模型在第7、28和42天时显示出显著更大的血管扩张，如最大直径倍数变化所反映的（以mm测量）分别为1.69 ± 0.07、1.99 ± 0.14和2.13 ± 0.09，而CaCl 2模型中分别为1.22 ± 0.04、1.48 ± 0.07和1.68±0.06（n=6; P<0.05）。第7天时弹性蛋白纤维完整性的半定量分级分析显示，与CaCl 2模型相比，CaPO 4模型中弹性蛋白降解显著增加（2.7±0.2对1.5±0.2，p<0.05，n=6）。在CaPO 4模型中发生了显著更高水平的细胞凋亡（术后1、2和3天的细胞凋亡指数：CaPO 4模型为0.26 ± 0.14、0.37± 0.14和0.33 ± 0.08，常规CaCl 2模型为0.012 ± 0.10、0.15± 0.02和0.12 ± 0.05）（n=3; p<0.05）。在第3天和第7天，在CaPO 4中还观察到巨噬细胞浸润和钙化的增强。在体外，与CaCl 2的混合物相比，CaPO 4诱导的VSMC凋亡约为3.7倍（n=4; p<0.0001）。我们的数据表明，CaPO 4模型加速动脉瘤的形成，细胞凋亡，巨噬细胞浸润和钙沉积的增强。这种改进的模型，其快速和强大的膨胀，可以作为一个新的模型AAA。
The Calcium Chloride (CaCl2) model is a widely accepted rodent model for abdominal aortic aneurysm (AAA). Calcium deposition, mainly consisting of calcium phosphate (CaPO4) crystals, has been reported to exist in both human and experimental aneurysms. CaPO4 crystal has been utilized for in vitro DNA transfection by mixing CaCl2 and Phosphate Buffered Saline (PBS). Here, we describe accelerated aneurysm formation resulting from a modification of the CaCl2 model. The modified CaCl2, the CaPO4 model, was created by applying PBS onto the mouse infrarenal aorta after CaCl2 treatment. Morphological, histological and immunohistochemical analyses were performed on arteries treated with both the CaPO4 model and the conventional CaCl2 model as control. In vitro methods were carried out using a mixture of CaCl2 and PBS to create CaPO4 crystals. CaPO4 induced apoptosis of primary cultured mouse vascular smooth muscle cells (VSMCs) was measured by DNA fragmentation ELISA. First, we showed that the CaPO4 model produces AAA, defined as an increase of 50% or greater in the diameter of the aorta; faster than in the CaCl2 model. CaPO4 model showed significantly larger aneurysmal dilation at 7, 28, and 42 days as reflected by a maximum diameter fold change (measured in mm) of 1.69 ± 0.07, 1.99 ± 0.14 and 2.13 ± 0.09 as opposed to 1.22 ± 0.04, 1.48 ± 0.07 and 1.68±0.06 as seen in CaCl2 model, respectively (n=6; P<0.05). A semi-quantitative grading analysis of elastin fiber integrity at 7 days revealed a significant increase in elastin degradation in the CaPO4 model as compared to CaCl2 model (2.7±0.2 vs 1.5±0.2, p<0.05, n=6). Significantly higher level of apoptosis occurred in the CaPO4 model (apoptosis index at 1, 2, and 3 days post-surgery: 0.26 ± 0.14, 0.37± 0.14, and 0.33 ± 0.08 for CaPO4 model and 0.012 ± 0.10, 0.15± 0.02, and 0.12 ± 0.05 for conventional CaCl2 model) (n=3; p<0.05). An enhancement of macrophage infiltration and calcification was also observed at 3 and 7 days in CaPO4. CaPO4 induced approximately 3.7 times more apoptosis in VSMCs when compared to a mixture of CaCl2 (n=4; p<0.0001) in vitro. Our data shows that the CaPO4 model accelerates aneurysm formation with the enhancement of apoptosis, macrophage infiltration and calcium deposition. This modified model, with its rapid and robust dilation, can be utilized as a new model for AAA.
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影响因子：
4.3
作者：
Kent, KC;Zwolak, RM;Cronenwett, JL
通讯作者：
Cronenwett, JL
影响因子：
15.9
作者：
Longo, GM;Xiong, WF;Baxter, BT
通讯作者：
Baxter, BT
影响因子：
13.3
作者：
Gao, Wentao;Ding, Wen-Xing;Yin, Xiao-Ming
通讯作者：
Yin, Xiao-Ming
影响因子：
15.9
作者：
GERTZ, SD;KURGAN, A;EISENBERG, D
通讯作者：
EISENBERG, D
影响因子：
4.4
作者：
Pazar, Borbala;Ea, Hang-Korng;Busso, Nathalie
通讯作者：
Busso, Nathalie